目的探讨沉默纤维蛋白原样蛋白1(fibrinogen-like protein 1,FGL1)对人肺腺癌PC-9细胞多西他赛敏感性的影响。方法Western blot法检测人正常支气管上皮细胞株BEAS-2B及人肺腺癌细胞株PC-9中的FGL1蛋白表达情况。以siRNA沉默PC-9细胞株中...目的探讨沉默纤维蛋白原样蛋白1(fibrinogen-like protein 1,FGL1)对人肺腺癌PC-9细胞多西他赛敏感性的影响。方法Western blot法检测人正常支气管上皮细胞株BEAS-2B及人肺腺癌细胞株PC-9中的FGL1蛋白表达情况。以siRNA沉默PC-9细胞株中FGL1基因,CCK-8法检测沉默FGL1对PC-9细胞增殖的抑制作用及对多西他赛敏感性的影响。结果与BEAS-2B细胞株比较,FGL1在PC9细胞株中呈高表达,其蛋白相对表达量是BEAS-2B细胞株的6.5倍,差异具有显著性(P<0.01)。通过转染FGL1siRNA沉默FGL1可增强多西他赛对PC-9细胞增殖的抑制作用。与FGL1siNC组细胞相比,FGL1siRNA组PC-9细胞的IC50值明显降低,差异具有显著性(P<0.01)。结论特异性沉默FGL1基因可抑制人肺腺癌细胞PC-9中FGL1的表达,抑制PC-9细胞的增殖,提高对多西他赛的敏感性。展开更多
Background Lumiracoxib is a highly selective cyclooxygenase-2 （COX-2） inhibitor with antiinflammatory, analgesic and antipyretic activities comparable with class specific drugs, but with much improved gastrointestin...Background Lumiracoxib is a highly selective cyclooxygenase-2 （COX-2） inhibitor with antiinflammatory, analgesic and antipyretic activities comparable with class specific drugs, but with much improved gastrointestinal safety. No studies have examined lumiracoxib for antitumorigenic activity on human nonsmall cell lung cancer cell lines in vitro or its possible molecular mechanisms. Methods The antiproliferative effect of lumiracoxib alone or combined with docetaxol on A549 and NCI-H460 lines was assessed by 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide （MTT） assay. Drug-drug interactions were analyzed using the coefficient of drug interaction （CDI） to characterize the interactions as synergism, additivity or antagonism. Morphological changes were observed by acridine orange fluorescent staining. Extent of apoptosis was determined by flow cytometry. Results Lumiracoxib （15-240 pmol/L） has an inhibitory effect on the proliferation of A549 and NCI-H460 cell lines in concentration- and time-dependent manners with the ICso values of 2597 pmol/L and 833 pmol/L, respectively. The synergistic effect was prominent when lumiracoxib （15-240 pmoVL） was combined with docetaxol （0.2-2 pmol/L） （CDI 〈1）. Fluorescent staining showed that lumiracoxib could induce apoptosis in A549 and NCI-H460 cells. Lumiracoxib treatment also caused an increase of the sub-G1 fraction in each cell line and resulted in an increase of G0/Gl-phase cells and a decrease of S-phase cells. Conclusions Lumiracoxib had antiproliferative effect on the human nonsmall cell lung cancer cell lines A549 and NCI-H460 and had a significant synergy with docetaxol, which may be related to apoptotic induction and cell cycle arrest.展开更多
文摘目的探讨长链非编码RNA(long non-coding RNA,LncRNA)牛磺酸上调基因1(taurine up-regulated gene 1,TUG1)对非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞生物学行为的影响及分子机制。方法采用qRT-PCR、Western blot分别检测组织(NSCLC癌组织、癌旁组织)和细胞(人正常气管上皮细胞HBE及NSCLC细胞A549、H2009、H1975)中TUG1、miR-142-5p及PD-L1的表达。将pcDNA-TUG1、si-TUG1、miR-142-5p mimics、si-TUG1+anti-miR-142-5p分别转染于A549,qRT-PCR、Western blot分别检测细胞中TUG1、miR-142-5p及PD-L1表达;CCK-8检测细胞增殖;Transwell检测细胞迁移、侵袭;双荧光素酶基因实验分别检测TUG1和miR-142-5p、miR-142-5p和PD-L1的关系;RNA免疫共沉淀检测miR-142-5p与TUG1的相互作用。结果NSCLC组织中TUG1(2.28±0.23 vs 1.00±0)、PD-L1(0.93±0.11 vs 0.25±0.01)表达高于癌旁组织,miR-142-5p(0.31±0.02 vs 1.00±0)表达低于癌旁组织(P<0.05);与人正常气管上皮细胞HBE比较,NSCLC细胞A549中TUG1(3.21±0.27 vs 1.00±0)、PD-L1(1.12±0.12 vs 0.24±0.01)表达水平最高,miR-142-5p(0.23±0.02 vs 1.00±0)表达水平最低(P<0.05);过表达TUG1促进A549细胞增殖、迁移、侵袭;下调TUG1或上调miR-142-5p表达均对A549细胞增殖、迁移、侵袭能力发挥抑制作用;TUG1负调控miR-142-5p,miR-142-5p负调控PD-L1;下调miR-142-5p减弱了沉默TUG1对A549细胞增殖、迁移及侵袭的抑制作用。结论TUG1在NSCLC组织和细胞中高表达,沉默TUG1通过miR-142-5p/PD-L1轴抑制A549细胞的增殖、迁移与侵袭。
文摘目的探讨沉默纤维蛋白原样蛋白1(fibrinogen-like protein 1,FGL1)对人肺腺癌PC-9细胞多西他赛敏感性的影响。方法Western blot法检测人正常支气管上皮细胞株BEAS-2B及人肺腺癌细胞株PC-9中的FGL1蛋白表达情况。以siRNA沉默PC-9细胞株中FGL1基因,CCK-8法检测沉默FGL1对PC-9细胞增殖的抑制作用及对多西他赛敏感性的影响。结果与BEAS-2B细胞株比较,FGL1在PC9细胞株中呈高表达,其蛋白相对表达量是BEAS-2B细胞株的6.5倍,差异具有显著性(P<0.01)。通过转染FGL1siRNA沉默FGL1可增强多西他赛对PC-9细胞增殖的抑制作用。与FGL1siNC组细胞相比,FGL1siRNA组PC-9细胞的IC50值明显降低,差异具有显著性(P<0.01)。结论特异性沉默FGL1基因可抑制人肺腺癌细胞PC-9中FGL1的表达,抑制PC-9细胞的增殖,提高对多西他赛的敏感性。
基金This work was supported by grants from the Natural Science Foundation of Anhui Province （No. 07021008） and the General Project of the Natural Science Foundation of the Department of Education, Anhui Province （No. KJ2007B142）.
文摘Background Lumiracoxib is a highly selective cyclooxygenase-2 （COX-2） inhibitor with antiinflammatory, analgesic and antipyretic activities comparable with class specific drugs, but with much improved gastrointestinal safety. No studies have examined lumiracoxib for antitumorigenic activity on human nonsmall cell lung cancer cell lines in vitro or its possible molecular mechanisms. Methods The antiproliferative effect of lumiracoxib alone or combined with docetaxol on A549 and NCI-H460 lines was assessed by 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide （MTT） assay. Drug-drug interactions were analyzed using the coefficient of drug interaction （CDI） to characterize the interactions as synergism, additivity or antagonism. Morphological changes were observed by acridine orange fluorescent staining. Extent of apoptosis was determined by flow cytometry. Results Lumiracoxib （15-240 pmol/L） has an inhibitory effect on the proliferation of A549 and NCI-H460 cell lines in concentration- and time-dependent manners with the ICso values of 2597 pmol/L and 833 pmol/L, respectively. The synergistic effect was prominent when lumiracoxib （15-240 pmoVL） was combined with docetaxol （0.2-2 pmol/L） （CDI 〈1）. Fluorescent staining showed that lumiracoxib could induce apoptosis in A549 and NCI-H460 cells. Lumiracoxib treatment also caused an increase of the sub-G1 fraction in each cell line and resulted in an increase of G0/Gl-phase cells and a decrease of S-phase cells. Conclusions Lumiracoxib had antiproliferative effect on the human nonsmall cell lung cancer cell lines A549 and NCI-H460 and had a significant synergy with docetaxol, which may be related to apoptotic induction and cell cycle arrest.
